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      Alexa Fluor™ 647 Mouse Anti-MR1
      Alexa Fluor™ 647 Mouse Anti-MR1

      Flow cytometric analysis of MR1 expression on Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMCs) were either cultured (16 h) with Acetyl-6-Formylpterin (+ Ac-6-FP; 2.3 µg/ml (10 µM); Cayman Chemicals Cat. No. 23303) or without Ac-6-FP (No Ac-6-FP) as indicated. The cells were then preincubated with Human BD Fc Block™ (Cat. No. 564219/564220) and stained with either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 565571; dashed line histograms) or Alexa Fluor™ 647 Mouse Anti-MR1 antibody (Cat. No. 568669/568670; solid line histograms) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histograms showing MR1 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

      Alexa Fluor™ 647 Mouse Anti-MR1

      Flow cytometric analysis of MR1 expression on B16-F0 cells. Cells from the Mouse B16-F0 (Skin Melanoma, ATCC® CRL-6322™) cell line were either cultured (16 h) with Acetyl-6-Formylpterin (+ Ac-6-FP; 2.3 µg/ml (10 µM); Cayman Chemicals Cat. No. 23303) or without Ac-6-FP (No Ac-6-FP) as indicated. The cells were then preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142] and stained with either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 565571; dotted line histograms) or Alexa Fluor™ 647 Mouse Anti-MR1 antibody (Cat. No. 568669/568670; solid line histograms) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histograms showing MR1 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

      Alexa Fluor™ 647 Mouse Anti-MR1 Alexa Fluor™ 647 Mouse Anti-MR1

      Flow cytometric analysis of MR1 expression on Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMCs) were either cultured (16 h) with Acetyl-6-Formylpterin (+ Ac-6-FP; 2.3 µg/ml (10 µM); Cayman Chemicals Cat. No. 23303) or without Ac-6-FP (No Ac-6-FP) as indicated. The cells were then preincubated with Human BD Fc Block™ (Cat. No. 564219/564220) and stained with either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 565571; dashed line histograms) or Alexa Fluor™ 647 Mouse Anti-MR1 antibody (Cat. No. 568669/568670; solid line histograms) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histograms showing MR1 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

      Flow cytometric analysis of MR1 expression on B16-F0 cells. Cells from the Mouse B16-F0 (Skin Melanoma, ATCC® CRL-6322™) cell line were either cultured (16 h) with Acetyl-6-Formylpterin (+ Ac-6-FP; 2.3 µg/ml (10 µM); Cayman Chemicals Cat. No. 23303) or without Ac-6-FP (No Ac-6-FP) as indicated. The cells were then preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142] and stained with either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 565571; dotted line histograms) or Alexa Fluor™ 647 Mouse Anti-MR1 antibody (Cat. No. 568669/568670; solid line histograms) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histograms showing MR1 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

      Product Details
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      BD Pharmingen™
      HLALS; MHC class I-like antigen MR-1; MHC class-I related-gene protein
      Human (QC Testing), Mouse (Tested in Development)
      Mouse IgG1, κ
      MR1 Extracellular Domain Complexed with β2m
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      AB_3684452
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.
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      Recommended Assay Procedures

      BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
      5. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
      8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
      11. For U.S. patents that may apply, see bd.com/patents.
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      568669 Rev. 2
      Antibody Details
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      8F2.F9

      The 8F2.F9 monoclonal antibody specifically recognizes MHC-related protein 1 (MR1). MRI is a nonclassical MHC class Ib molecule. It is comprised of a ~40 kDa, highly conserved transmembrane a heavy chain that is a type I glycoprotein which is noncovalently-associated with an invariant ß2-microglobulin (ß2m) light chain. The N-terminal extracellular region of the HLA class I heavy chain is comprised of three domains (a1, a2, and a3). The a1 and a2 domains form a closed antigen-binding groove that accommodates small antigens whereas the a3 domain interacts with ß2m. MR1 is an antigen-presenting molecule that is specialized in displaying metabolites derived from microbial riboflavin biosynthesis to a small population of aß T-cells expressing an invariant TCR a chain called mucosal-associated invariant T-cells (MAIT). This function is essential to the development and expansion of MAIT cells. The 8F2.F9 and 26.5 monoclonal antibodies reportedly bind to different MRI epitopes.

      568669 Rev. 2
      Format Details
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      Alexa Fluor™ 647
      Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      Alexa Fluor™ 647
      Red 627-640 nm
      653 nm
      669 nm
      568669 Rev.2
      Citations & References
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      View product citations for antibody "568669" on CiteAb

      Development References (8)

      1. Abós B, Gómez Del Moral M, Gozalbo-López B, López-Relaño J, Viana V, Martínez-Naves E. Human MR1 expression on the cell surface is acid sensitive, proteasome independent and increases after culturing at 26°C.. Biochem Biophys Res Commun. 2011; 411(3):632-6. (Biology: Flow cytometry). View Reference
      2. Corbett AJ, Awad W, Wang H, Chen Z. Antigen Recognition by MR1-Reactive T Cells; MAIT Cells, Metabolites, and Remaining Mysteries.. Front Immunol. 2020; 11:1961. (Biology). View Reference
      3. Huang S, Gilfillan S, Cella M, et al. Evidence for MR1 antigen presentation to mucosal-associated invariant T cells.. J Biol Chem. 2005; 280(22):21183-93. (Immunogen: Flow cytometry). View Reference
      4. Lamichhane R, Ussher JE. Expression and trafficking of MR1.. Immunology. 2017; 151(3):270-279. (Biology). View Reference
      5. McWilliam HE, Eckle SB, Theodossis A, et al. The intracellular pathway for the presentation of vitamin B-related antigens by the antigen-presenting molecule MR1.. Nat Immunol. 2016; 17(5):531-7. (Clone-specific: Flow cytometry). View Reference
      6. Miley MJ, Truscott SM, Yu YY, et al. Biochemical features of the MHC-related protein 1 consistent with an immunological function.. J Immunol. 2003; 170(12):6090-8. (Biology: Blocking, Flow cytometry, In vivo exacerbation). View Reference
      7. Salio M, Awad W, Veerapen N, et al. Ligand-dependent downregulation of MR1 cell surface expression.. Proc Natl Acad Sci U S A. 2020; 117(19):10465-10475. (Clone-specific: Flow cytometry). View Reference
      8. Yan J, Allen S, McDonald E, et al. MAIT Cells Promote Tumor Initiation, Growth, and Metastases via Tumor MR1.. Cancer Discov. 2020; 10(1):124-141. (Clone-specific). View Reference
      View All (8) View Less
      568669 Rev. 2

      Please refer to Support Documents for Quality Certificates

       

      Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

       

      Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

      For Research Use Only. Not for use in diagnostic or therapeutic procedures.